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3D multiplexed immunoplasmonics microscopy.

Éric Bergeron1, Sergiy Patskovsky, David Rioux

  • 1Laser Processing and Plasmonics Laboratory, Department of Engineering Physics, Polytechnique Montréal, C.P. 6079, Succursale Centre-Ville, Montréal, QC H3C 3A7, Canada. michel.meunier@polymtl.ca.

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Summary
This summary is machine-generated.

Immunoplasmonics microscopy uses nanoparticle-labeled antibodies for precise cell surface biomarker detection. This novel technique offers a photostable, cost-efficient alternative to immunofluorescence for clinical applications.

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Area of Science:

  • Biomedical Engineering
  • Nanotechnology
  • Microscopy

Background:

  • Cell surface biomarkers are crucial for diagnosing diseases and guiding treatment.
  • Immunofluorescence is the current standard but suffers from photobleaching and spectral limitations.
  • A need exists for more stable and multiplexed biomarker detection methods.

Purpose of the Study:

  • To demonstrate accurate multiplexed detection of cell surface biomarkers using immunoplasmonics.
  • To develop a 3D localization and spectral identification method for immunoplasmonic labels.
  • To establish immunoplasmonics as a viable clinical alternative to immunofluorescence.

Main Methods:

  • Functionalized plasmonic nanoparticles (fNPs) targeting CD44, EGFR, and KV1.1 biomarkers were synthesized.
  • Selective labeling of biomarkers on human cancer cells (MDA-MB-231) and reference cells (661W) was performed.
  • Reflected light microscopy (RLM) with fast z-scanning and spectral filters was used for 3D localization and identification of fNPs.

Main Results:

  • Accurate multiplexed detection of three cell surface biomarkers was achieved using fNPs (AgNSs, AuNSs, AuNRs).
  • Labeling efficiency was comparable to immunofluorescence.
  • A novel 3D localization and spectral identification method was successfully implemented.

Conclusions:

  • 3D multiplexed immunoplasmonics microscopy enables precise, stable, and multiplexed detection of cell surface biomarkers.
  • The technology is compatible with standard microscopy equipment, making it suitable for clinical settings.
  • Immunoplasmonics offers a cost-efficient and advanced alternative to immunofluorescence for clinical diagnostics.